nogrЭкспериментальная и клиническая гастроэнтерологияExperimental and Clinical Gastroenterology1682-8658«Global Media Technologies»10.31146/1682-8658-ecg-218-10-160-167nogr-2560Research ArticleГЕПАТОЛОГИЯHEPATOLOGYУровень экспрессии длинных некодирующих РНК MALAT1, GAS5, DANCR и TUG1 в лейкоцитах периферической крови пациентов с неалкогольной жировой болезнью печениExpression level of long non-coding RNA MALAT1, GAS5, DANCR and TUG1 in peripheral blood leukocytes of patients with non-alcoholic fatty liver diseasehttps://orcid.org/0000-0001-7620-7065КурбатоваИ. В.KurbatovaI. V.irina7m@yandex.ruhttps://orcid.org/0000-0003-3849-6796ВасильеваА. В.VasilevaA. V.noemail@neicon.ruhttps://orcid.org/0000-0001-8697-2086ТопчиеваЛ. В.TopchievaL. V.noemail@neicon.ruhttps://orcid.org/0000-0003-2613-5694ДудановаО. П.DudanovaO. P.noemail@neicon.ruhttps://orcid.org/0000-0003-3830-6446ШиповскаяА. А.ShopovskayaA. A.noemail@neicon.ruИнститут биологии - обособленное подразделение Федерального государственного бюджетного учреждения науки Федерального исследовательского центра «Карельский научный центр Российской академии наук»РоссияInstitute of Biology of the Karelian Research Centre of the Russian Academy of Sciences (IB KarRC RAS)Russian FederationФедеральное государственное бюджетное образовательное учреждение высшего образования «Петрозаводский государственный университет»РоссияPetrozavodsk State UniversityRussian Federation202315032024010160167Copyright © Курбатова И.В., Васильева А.В., Топчиева Л.В., Дуданова О.П., Шиповская А.А., 20242024Курбатова И.В., Васильева А.В., Топчиева Л.В., Дуданова О.П., Шиповская А.А.Kurbatova I.V., Vasileva A.V., Topchieva L.V., Dudanova O.P., Shopovskaya A.A.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.nogr.org/jour/article/view/2560

Цель: Сравнительный анализ уровня экспрессии длинных некодирующих РНК MALAT1, GAS5, DANCR, TUG1 в лейкоцитах периферической крови (ЛПК) здоровых людей и пациентов с НАЖБП (стеатозом печени, неалкогольным стеатогепатитом (НАСГ) разной активности, циррозом печени). Материалы и методы: Обследовано 106 больных с диагнозом НАЖБП, установленным впервые: 31 пациент со стеатозом печени (СП), 64 пациента с НАСГ слабой (СА), умеренной (УА) и высокой (ВА) активности и 11 пациентов на стадии цирроза печени (ЦП). Контрольную группу составили 30 здоровых доноров. Уровень мРНК генов TUG1, DANCR, MALAT1, GAS5 в ЛПК определяли методом ПЦР-РВ. Результаты: Зарегистрирован более высокий уровень экспрессии гена TUG1 в ЛПК больных НАСГ-СА по сравнению с СП, а также выявлена тенденция повышения уровня мРНК TUG1 в ЛПК при нарастании активности НАСГ, что указывает на возможность использования уровня экспрессии TUG1 в ЛПК в качестве малоинвазивного диагностического (для разграничения СП и НАСГ-СА) и прогностического маркера (при прогрессировании НАЖБП). Анализ уровня экспрессии днРНК MALAT1 показал отсутствие достоверных различий между всеми исследованными группами. Получены результаты, свидетельствующие о сложной динамике уровня экспрессии GAS5: уровень транскриптов повышается при формировании стеатоза печени и затем снижается при переходе в НАСГ. Показано, что уровень экспрессии DANCR в ЛПК пациентов с НАСГ-СА ниже, чем у пациентов со стеатозом печени и НАСГ-УА. Заключение: Получены новые данные по уровню экспрессии днРНК MALAT1, GAS5, DANCR, TUG1 в ЛПК пациентов с НАЖБП, свидетельствующие о возможности использования уровня экспрессии TUG1 в ЛПК в качестве малоинвазивного диагностического и прогностического маркера при НАЖБП. Также показано, что уровень мРНК DANCR в ЛПК может иметь определенную диагностическую ценность при разграничении СП и НАСГ-СА.

Purpose: Comparative analysis of the expression level of long non-coding RNAs MALAT1, GAS5, DANCR, TUG1 in peripheral blood leukocytes (PBL) of healthy people and patients with NAFLD (liver steatosis, NASH of varying activity, liver cirrhosis). Materials and methods: We examined 106 patients diagnosed with NAFLD for the first time: 31 patients with liver steatosis (LS), 64 patients with weak (WA), moderate (MA) and high (HA) NASH activity and 11 patients at the stage of liver cirrhosis (LC). The control group consisted of 30 healthy donors. The mRNA level of the TUG1, DANCR, MALAT1, GAS5 genes in PBL was determined by RT-PCR. Results: A higher level of expression of the TUG1 gene was registered in the PBL of patients with NASH-WA compared to LS, and a tendency was revealed to increase the level of TUG1 mRNA in the PBL with increasing NASH activity, which indicates the possibility of using the level of TUG1 expression in the PBL as a minimally invasive diagnostic (to distinguish between LS and NASH-WA) and a prognostic marker (with the progression of NAFLD). Analysis of the expression level of lncRNA MALAT1 showed no significant differences between all studied groups. Results were obtained indicating complex dynamics of the GAS5 expression level: the level of transcripts increases during the formation of liver steatosis and then decreases during the transition to NASH. It was shown that the level of DANCR expression in the PBL of patients with NASH-WA is significantly lower than in patients with liver steatosis and NASH-MA. Conclusion: New data were obtained on the expression level of the MALAT1, GAS5, DANCR, TUG1 lncRNAs in the PBL of patients with NAFLD, indicating the possibility of using the level of TUG1 expression in the PBL as a minimally invasive diagnostic and prognostic marker in NAFLD. It has also been shown that the level of DANCR mRNA in PBL may have some diagnostic value in distinguishing between LS and NASH-WA.

неалкогольная жировая болезнь печени (НАЖБП)неалкогольный стеатогепатит (НАСГ)днРНКMALAT1GAS5DANCRTUG1non-alcoholic fatty liver disease (NAFLD)non-alcoholic steatohepatitis (NASH)lncRNAMALAT1GAS5DANCRTUG1ReferencesLazebnik L. B., Golovanova E. V., Turkina S. V. et al. Non-alcoholic fatty liver disease in adults: clinic, diagnostics, treatment. Guidelines for therapists, third version. Experimental and Clinical Gastroenterology. 2021;1(1):4-52. (In Russ.) doi: 10.31146/1682-8658-ecg-185-1-4-52.@@ Лазебник Л. Б., Голованова Е. В., Туркина С. В. и др. Неалкогольная жировая болезнь печени у взрослых: клиника, диагностика, лечение. Рекомендации для терапевтов, третья версия. Экспериментальная и клиническая гастроэнтерология. 2021; 185(1): 4-52. doi: 10.31146/1682-8658-ecg-185-1-4-52.Lazebnik L. B., Golovanova E. V., Turkina S. V. et al. Non-alcoholic fatty liver disease in adults: clinic, diagnostics, treatment. Guidelines for therapists, third version. Experimental and Clinical Gastroenterology. 2021;1(1):4-52. (In Russ.) doi: 10.31146/1682-8658-ecg-185-1-4-52.@@ Лазебник Л. Б., Голованова Е. В., Туркина С. В. и др. Неалкогольная жировая болезнь печени у взрослых: клиника, диагностика, лечение. Рекомендации для терапевтов, третья версия. Экспериментальная и клиническая гастроэнтерология. 2021; 185(1): 4-52. doi: 10.31146/1682-8658-ecg-185-1-4-52.Ye L., Zhao D., Xu Y. et.al. LncRNA-Gm9795 promotes inflammation in non-alcoholic steatohepatitis via NF-κB/JNK pathway by endoplasmic reticulum stress. J. Transl. Med. 2021; 19:101. doi: 10.1186/s12967-021-02769-7.Ye L., Zhao D., Xu Y. et.al. LncRNA-Gm9795 promotes inflammation in non-alcoholic steatohepatitis via NF-κB/JNK pathway by endoplasmic reticulum stress. J. Transl. Med. 2021; 19:101. doi: 10.1186/s12967-021-02769-7.Leti F., Legendre C., Still C. D. et al. Altered expression of MALAT1 lncRNA in nonalcoholic steatohepatitis fibrosis regulates CXCL5 in hepatic stellate cells. Translational Research. 2017; 190: 25-39. doi: 10.1016/j.trsl.2017.09.001.Leti F., Legendre C., Still C. D. et al. Altered expression of MALAT1 lncRNA in nonalcoholic steatohepatitis fibrosis regulates CXCL5 in hepatic stellate cells. Translational Research. 2017; 190: 25-39. doi: 10.1016/j.trsl.2017.09.001.Bu F., Wang A., Zhu Y. et al. LncRNA NEAT1: shedding light on mechanisms and opportunites in liver diseases. Liver Int. 2020; 40(11): 2612-2626. doi: 10.1111/liv.14629.Bu F., Wang A., Zhu Y. et al. LncRNA NEAT1: shedding light on mechanisms and opportunites in liver diseases. Liver Int. 2020; 40(11): 2612-2626. doi: 10.1111/liv.14629.He Z., Yang D., Fan X. et al. The Roles and Mechanisms of lncRNAs in Liver Fibrosis.Int. J. Mol. Sci. 2020; 21(4): 1482. doi:10.3390/ijms21041482.He Z., Yang D., Fan X. et al. The Roles and Mechanisms of lncRNAs in Liver Fibrosis.Int. J. Mol. Sci. 2020; 21(4): 1482. doi:10.3390/ijms21041482.Rohilla S., Kaur S., Puria R. Long non-coding RNA in Non-alcoholic fatty liver disease. Adv Clin Chem. 2022;110:1-35. doi: 10.1016/bs.acc.2022.06.001.Rohilla S., Kaur S., Puria R. Long non-coding RNA in Non-alcoholic fatty liver disease. Adv Clin Chem. 2022;110:1-35. doi: 10.1016/bs.acc.2022.06.001.Zeng Q., Liu C. H., Wu D. et al. LncRNA and circRNA in Patients with Non-Alcoholic Fatty Liver Disease: A Systematic Review. Biomolecules 2023; 13, 560. doi:10.3390/biom13030560.Zeng Q., Liu C. H., Wu D. et al. LncRNA and circRNA in Patients with Non-Alcoholic Fatty Liver Disease: A Systematic Review. Biomolecules 2023; 13, 560. doi:10.3390/biom13030560.Yu F., Zheng J., Mao Y. et al. Long Non-coding RNA Growth Arrest-specific Transcript 5 (GAS5) Inhibits Liver Fibrogenesis through a Mechanism of Competing Endogenous RNA. J Biol Chem. 2015; 290(47): 28286-28298. doi: 10.1074/jbc.M115.683813.Yu F., Zheng J., Mao Y. et al. Long Non-coding RNA Growth Arrest-specific Transcript 5 (GAS5) Inhibits Liver Fibrogenesis through a Mechanism of Competing Endogenous RNA. J Biol Chem. 2015; 290(47): 28286-28298. doi: 10.1074/jbc.M115.683813.Han M.-H., Lee J. H., Kim G. et al. Expression of the Long Noncoding RNA GAS5 Correlates with Liver Fibrosis in Patients with Nonalcoholic Fatty Liver Disease. Genes. 2020; 11, 545. doi: 10.3390/genes11050545.Han M.-H., Lee J. H., Kim G. et al. Expression of the Long Noncoding RNA GAS5 Correlates with Liver Fibrosis in Patients with Nonalcoholic Fatty Liver Disease. Genes. 2020; 11, 545. doi: 10.3390/genes11050545.Mukherjee A. G., Wanjari U. R., Gopalakrishnan A. V. et al. Exploring the Regulatory Role of ncRNA in NAFLD: A Particular Focus on PPARs. Cells. 2022; 11, 3959. doi: 10.3390/cells11243959.Mukherjee A. G., Wanjari U. R., Gopalakrishnan A. V. et al. Exploring the Regulatory Role of ncRNA in NAFLD: A Particular Focus on PPARs. Cells. 2022; 11, 3959. doi: 10.3390/cells11243959.Tripathi V., Ellis J. D., Shen Z. et al. The nuclear-retained noncoding RNA MALAT1 regulates alternative splicing by modulating SR splicing factor phosphorylation. Mol Cell. 2010; 39(6): 925-38. doi: 10.1016/j.molcel.2010.08.011.Tripathi V., Ellis J. D., Shen Z. et al. The nuclear-retained noncoding RNA MALAT1 regulates alternative splicing by modulating SR splicing factor phosphorylation. Mol Cell. 2010; 39(6): 925-38. doi: 10.1016/j.molcel.2010.08.011.Lu J., Guo J., Liu J. et al. Long non-coding RNA MALAT1: A key player in liver diseases. Frontiers in Medicine. 2022; 734643. doi: 10.3389/fmed.2021.734643.Lu J., Guo J., Liu J. et al. Long non-coding RNA MALAT1: A key player in liver diseases. Frontiers in Medicine. 2022; 734643. doi: 10.3389/fmed.2021.734643.Cui J., Wang Y., Xue H. Long non-coding RNA GAS5 contributes to the progression of nonalcoholic fatty liver disease by targeting the microRNA-29a-3p/NOTCH2 axis. Bioengineered. 2022; 13(4): 8370-8381. doi: 10.1080/21655979.2022.2026858.Cui J., Wang Y., Xue H. Long non-coding RNA GAS5 contributes to the progression of nonalcoholic fatty liver disease by targeting the microRNA-29a-3p/NOTCH2 axis. Bioengineered. 2022; 13(4): 8370-8381. doi: 10.1080/21655979.2022.2026858.Chen, T., Meng, Y., Zhou, Z. et al. GAS5 protects against nonalcoholic fatty liver disease via miR-28a-5p/MARCH7/NLRP3 axis-mediated pyroptosis. Cell Death Differ. 2023; 30: 1829-1848. doi: 10.1038/s41418-023-01183-4.Chen, T., Meng, Y., Zhou, Z. et al. GAS5 protects against nonalcoholic fatty liver disease via miR-28a-5p/MARCH7/NLRP3 axis-mediated pyroptosis. Cell Death Differ. 2023; 30: 1829-1848. doi: 10.1038/s41418-023-01183-4.Lin Y. H., Wu M. H., Huang Y. H. et al. Taurine up-regulated gene 1 functions as a master regulator to coordinate glycolysis and metastasis in hepatocellular carcinoma. Hepatology. 2018; 67(1): 188-203. doi: 10.1002/hep.29462.Lin Y. H., Wu M. H., Huang Y. H. et al. Taurine up-regulated gene 1 functions as a master regulator to coordinate glycolysis and metastasis in hepatocellular carcinoma. Hepatology. 2018; 67(1): 188-203. doi: 10.1002/hep.29462.Guo D., Li Y., Chen Y. et al. DANCR promotes HCC progression and regulates EMT by sponging miR-27a-3p via ROCK1/LIMK1/COFILIN1 pathway. Cell Prolif. 2019; 52(4): e12628. doi: 10.1111/cpr.12628.Guo D., Li Y., Chen Y. et al. DANCR promotes HCC progression and regulates EMT by sponging miR-27a-3p via ROCK1/LIMK1/COFILIN1 pathway. Cell Prolif. 2019; 52(4): e12628. doi: 10.1111/cpr.12628.Pinto J. P., Dias V., Zoller H. et.al. Hepcidin messenger RNA expression in human lymphocytes. Immunology. 2010; 130(2): 217-230. doi: 10.1111/j.1365-2567.2009.03226.x.Pinto J. P., Dias V., Zoller H. et.al. Hepcidin messenger RNA expression in human lymphocytes. Immunology. 2010; 130(2): 217-230. doi: 10.1111/j.1365-2567.2009.03226.x.Wang W. Y., Wang Y. F., Ma P. et al. Taurine-upregulated gene 1: a vital long non-coding RNA associated with cancer in humans (review). Mol Med Rep. 2017; 16(5): 6467-6471. doi: 10.3892/mmr.2017.7472.Wang W. Y., Wang Y. F., Ma P. et al. Taurine-upregulated gene 1: a vital long non-coding RNA associated with cancer in humans (review). Mol Med Rep. 2017; 16(5): 6467-6471. doi: 10.3892/mmr.2017.7472.Han X., Hong Y., Zhang K. Tug1 is involved in liver fibrosis and activation of hscs by regulating mir-29b. Biochem. Biophys. Res.Commun. 2018; 503: 1394-1400. doi: 10.1016/j.bbrc.2018.07.054.Han X., Hong Y., Zhang K. Tug1 is involved in liver fibrosis and activation of hscs by regulating mir-29b. Biochem. Biophys. Res.Commun. 2018; 503: 1394-1400. doi: 10.1016/j.bbrc.2018.07.054.Qin C. F., Zhao F. L. Long non-coding RNA TUG1 can promote proliferation and migration of pancreatic cancer via EMT pathway. Eur Rev Med Pharmacol Sci. 2017; 21(10): 2377-2384. PMID: 28617552.Qin C. F., Zhao F. L. Long non-coding RNA TUG1 can promote proliferation and migration of pancreatic cancer via EMT pathway. Eur Rev Med Pharmacol Sci. 2017; 21(10): 2377-2384. PMID: 28617552.Sun J., Ding C., Yang Z., et al. The long non-coding RNA TUG1 indicates a poor prognosis for colorectal cancer and promotes metastasis by affecting epithelial-mesenchymal transition. J Transl Med. 2016; 14:42. doi: 10.1186/s12967-016-0786-z.Sun J., Ding C., Yang Z., et al. The long non-coding RNA TUG1 indicates a poor prognosis for colorectal cancer and promotes metastasis by affecting epithelial-mesenchymal transition. J Transl Med. 2016; 14:42. doi: 10.1186/s12967-016-0786-z.Hu M., Wang Y., Meng Y. et al. Hypoxia induced-disruption of lncRNA TUG1/PRC2 interaction impairs human trophoblast invasion through epigenetically activating Nodal/ALK7 signalling. J Cell Mol Med. 2022; 26: 4087-4100. doi: 10.1111/jcmm.17450.Hu M., Wang Y., Meng Y. et al. Hypoxia induced-disruption of lncRNA TUG1/PRC2 interaction impairs human trophoblast invasion through epigenetically activating Nodal/ALK7 signalling. J Cell Mol Med. 2022; 26: 4087-4100. doi: 10.1111/jcmm.17450.Xiang J., Deng Y. Y., Liu H. X. et al. LncRNA MALAT1 promotes PPARα/CD36-mediated hepatic lipogenesis in nonalcoholic fatty liver disease by modulating miR-206/ARNT axis. Frontiers in Bioengineering and Biotechnology. 2022; 10: 858558. doi: 10.3389/fbioe.2022.858558.Xiang J., Deng Y. Y., Liu H. X. et al. LncRNA MALAT1 promotes PPARα/CD36-mediated hepatic lipogenesis in nonalcoholic fatty liver disease by modulating miR-206/ARNT axis. Frontiers in Bioengineering and Biotechnology. 2022; 10: 858558. doi: 10.3389/fbioe.2022.858558.Zeng Q., Liu C. H., Wu D. et al. LncRNA and circRNA in Patients with Non-Alcoholic Fatty Liver Disease: A Systematic Review. Biomolecules. 2023; 13(3): 560. doi: 10.3390/biom13030560.Zeng Q., Liu C. H., Wu D. et al. LncRNA and circRNA in Patients with Non-Alcoholic Fatty Liver Disease: A Systematic Review. Biomolecules. 2023; 13(3): 560. doi: 10.3390/biom13030560.Liao X., Chen J., Luo D. et al. Prognostic value of long non-coding RNA MALAT1 in hepatocellular carcinoma: A study based on multi-omics analysis and RT-PCR validation. Pathology and Oncology Research. 2023; 28: 1610808. doi: 10.3389/pore.2022.1610808.Liao X., Chen J., Luo D. et al. Prognostic value of long non-coding RNA MALAT1 in hepatocellular carcinoma: A study based on multi-omics analysis and RT-PCR validation. Pathology and Oncology Research. 2023; 28: 1610808. doi: 10.3389/pore.2022.1610808.Su S. B., Tao L., Liang X. L. et.al. Long noncoding RNA GAS5 inhibits LX-2 cells activation by suppressing NF-κB signalling through regulation of the miR-433-3p/TLR10 axis. Dig Liver Dis. 2022; 54(8): 1066-1075. doi: 10.1016/j.dld.2021.11.002.Su S. B., Tao L., Liang X. L. et.al. Long noncoding RNA GAS5 inhibits LX-2 cells activation by suppressing NF-κB signalling through regulation of the miR-433-3p/TLR10 axis. Dig Liver Dis. 2022; 54(8): 1066-1075. doi: 10.1016/j.dld.2021.11.002.Yuan S., Wang J., Yang F. et.al. Long noncoding RNA DANCR increases stemness features of hepatocellular carcinoma by derepression of CTNNB1. Hepatology. 2016; 63(2):499-511. doi: 10.1002/hep.27893.Yuan S., Wang J., Yang F. et.al. Long noncoding RNA DANCR increases stemness features of hepatocellular carcinoma by derepression of CTNNB1. Hepatology. 2016; 63(2):499-511. doi: 10.1002/hep.27893.Ghafouri-Fard S., Khoshbakht T., Hussen B. M. et.al. A review on the role of DANCR in the carcinogenesis. Cancer Cell Int. 2022; 22(1): 194. doi: 10.1186/s12935-022-02612-z.Ghafouri-Fard S., Khoshbakht T., Hussen B. M. et.al. A review on the role of DANCR in the carcinogenesis. Cancer Cell Int. 2022; 22(1): 194. doi: 10.1186/s12935-022-02612-z.Yang L., Jiang M. N., Liu Y. et.al. Crosstalk between lncRNA DANCR and miR-125b-5p in HCC cell progression. Tumori J. 2021;107(6): 504-513. doi: 10.1177/0300891620977010.Yang L., Jiang M. N., Liu Y. et.al. Crosstalk between lncRNA DANCR and miR-125b-5p in HCC cell progression. Tumori J. 2021;107(6): 504-513. doi: 10.1177/0300891620977010.

The authors declare that there are no conflicts of interest present.